Search results for: Streptococcus agalactiae

Authors: Natesan Balasubramanian, Palpandi Pounpandi, Venkatraman Thamil Priya, Vellasamy Shanmugaiah, Karubbiah Balakrishnan, Mandayam Anandam Thirunarayan

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Streptococcus agalactiae is commonly known as Group B Streptococcus (GBS) and it is the most common cause of life-threatening bacterial infection. GBS first considered as a veterinary pathogen causing mastitis in cattle later becomes a human pathogen for severe neonatal infections. In this present study, a total of 20 new clinical isolates of S. agalactiae were collected from male (6) and female patient (14) with different age group. The isolates were from Urinary tract infection (UTI), blood, pus and eye ulcer. All the 20 S. agalactiae isolates has clear hemolysis properties on blood agar medium and were identified by serogrouping and MALTI-TOF-MS analysis. Antibiotic susceptibility/resistance test was performed for 20 S. agalactiae isolates, further phenotypic resistance pattern was observed for tetracycline, vancomycin, ampicillin and penicillin. Genotypically we found two antibiotic resistance genes such as Betalactem antibiotic resistance gene (Tem) (70%) and tetracycline resistance gene Tet(O) 15% in our isolates. Six virulence factors encoding genes were performed by PCR in twenty GBS isolates, cfb gene (100%), followed by, cylE(90.47%), lmp(85.7%), bca(71.42%), rib (38%) and low frequency in bac gene (4.76%) were determined. Most of the S. agalactiae isolates produced strong biofilm in the polystyrene surface (hydrophobic), and low-level biofilm formation was found in glass tube (hydrophilic) surface. lytR is secreted protein and localized in bacterial cell wall, extra cellular membrane, and cytoplasm. In silico docking studies were performed for lytR protein with four antibiofilm compounds, including a peptide (PR39) with the docking study showed peptide has strong interaction followed by ellagic acid and interaction length is 2.95, 2.97 and 2.95 A°. In ligand EGCGO10 and O11 two atoms intract with lytR (Leu271), with binding bond affinity length is 3.24 and 3.14. The aminoacid Leu 271 is act as an impartant aminoacid, since ellagic acid and EGCG interact with same aminoacid.

Keywords: antibiotics, biofilms, clinical isolates, S. agalactiae, virulence

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Authors: Isabel Cristina Ortiz Trujillo, Lina Maria Martinez Sanchez, Felipe Hernández Restrepo, Daniel Gallego Gonzalez, Natalia Vargas Grisales, Camilo Andrés Agudelo Vélez

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Chorioamnionitis, is a pregnancy infection, causes different fetal and maternal symptoms. Streptococcus agalactiae present in the normal vaginal microflora of some women, favouring its abnormal multiplication during pregnancy, causing perinatal morbidity and mortality. Objective. Describe the clinical and epidemiological profile of the patients with diagnosis of clinical chorioanmionitis. Methodology. Descriptive, cross-sectional study. The population was patients with diagnosis of clinical chorioanmionitis. The information was taken from the medical records. The research was approved by the Ethics Committee. We used the program SPSS ® version 17.0 (SPSS Inc; Chicago, Illinois, USA) for the information analysis, descriptive statistics were used. Results. 78 patients in total with clinical chorioamnionitis, with a mean age of 26.3 ±5, 8 years old, the 69.2% primigravid women. 2.6% of women had positive culture for Streptococcus agalactiae in urine sample during current pregnancy and 30.7% had received some kind of antibiotics during current pregnancy. The 57.7% had 37 to 40 weeks of gestation in the current pregnancy it was calculated more frequently by ultrasound (66.7% in first quarter, 11.5% in the second and 1.9% in the third). In a 60.3% way of termination of pregnancy was vaginal and a 35.9 percent were caesarean section. Among the women in the study, a 30.8% had premature rupture of membranes. Conclusion. The chorioamnionitis continues to be an important cause of antibiotic use during pregnancy or labour and the decision to do a caesarean, with highest percentage in pregnancies-preterm and preterm premature rupture of membranes.

Keywords: chorioamnionitis, Streptococcus agalactiae, pregnancy complications, infectious

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Authors: Nastaran Hemmati, Farhad Nikkhahi, Amir Javadi, Sahar Eskandarion, Seyed Mahmuod Amin Marashi

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Neisseria meningitidis, Escherichia coli K, Streptococcus agalactiae, and Streptococcus pneumoniae cause 90% of bacterial meningitis. Almost all infected people die or have irreversible neurological complications. Therefore, it is essential to have a diagnostic kit with the ability to quickly detect these fatal infections. The project involved 212 patients from whom cerebrospinal fluid samples were obtained. After total genome extraction and performing multiplex quantitative polymerase chain reaction (qPCR), the presence or absence of each infectious factor was determined by comparing with standard strains. The specificity, sensitivity, positive predictive value, and negative predictive value calculated were 100%, 92.9%, 50%, and 100%, respectively. So, due to the high specificity and sensitivity of the designed primers, they can be used instead of bacterial culture that takes at least 24 to 48 hours. The remarkable benefit of this method is associated with the speed (up to 3 hours) at which the procedure could be completed. It is also worth noting that this method can reduce the personnel unintentional errors which may occur in the laboratory. On the other hand, as this method simultaneously identifies four common factors that cause bacterial meningitis, it could be used as an auxiliary method diagnostic technique in laboratories particularly in cases of emergency medicine.

Keywords: cerebrospinal fluid, meningitis, quantitative polymerase chain reaction, simultaneous detection, diagnosis testing

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Authors: Kavita Jaidiya, Anju Chahar, Chitra Jaidiya

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The present study was conducted on 200 quarters from 50 apparently healthy cows. Samples are subjected to California Mastitis Test (CMT), cultural examination, and mPCR. Milk samples were also subjected to changes in composition Viz. fat, protein, and lactose. The prevalence of subclinical mastitis based on culture examination was 30(60/200), 36 (72/200), and 40 percent (93/200) based on CMT, culture examination, and mPCR on a quarterly basis. The prevalence of subclinical mastitis on animal basis was 40 (20/50), 46 (23/50), and 52 percent (26/50) based on CMT, Culture examination, and mPCR. The highest prevalence was observed in IVth parity on a quarterly basis and in Vth parity on cow basis. On culture examination, Staphylococcus aureus was the most prevalent organism (50.56%), followed by Streptococcus dysaglactiae (11.33%), E. coli (7.8 %), Staphylococcus agalactiae (13.48 %), Staphylococcus epidermidis (2.2 %), Streptococcus hyicus (6.94%), Streptococcus uberis (5.16%), Klebsiella pneumonia (6.74%). On isolation by bacterial mPCR, Staphylococcus spp. (42%) was the major pathogen. Organisms isolated in mixed infections are Streptococcus spp., Klebsiella pneumonia, E.coli and Pseudomonas aeruginous. The average mean value of fat, protein, and lactose content in subclinically affected milk samples were 3.40 ± 0.101, 3.009 ± 0.033, and 4.48 ± 0.03, and the mean value of fat, protein, and lactose content in normal milk were 4.13 ± 0.035, 3.39 ± 0.021, and 5.10 ± 0.016. The mean blood level of reduced glutathione in subclinical mastitis (30.44 ± 1.87 ng/ml) was lower than healthy cows (47.98 ± 4.04ng/ml). The concentration of malondialdehyde (10.026 ± 0.21mmol/L) in subclinical mastitis was significantly higher as compared to healthy group cows (2.19 ± 0.23mmol/L).

Keywords: cow, subclinical mastitis, mPCR, California Mastitis test

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Authors: Sara Sousa, Veronique Gomes, Nélio Veiga, Maria José Correia

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Objectives: To assess the presence or absence of Streptococcus mutans, Streptococcus gordonii and Streptococcus salivarius in the oral biofilm of children in an elementary school of Viseu, Portugal, and verify the relationship between Streptococcus gordonii and Streptococcus salivarius and the absence of dental caries. Methods: A cross-sectional study was designed with a final sample of 40 children aged 6-11 years old. Oral examination was accomplished with the identification of their oral health status and oral biofilm collection. Analysis of biological samples by molecular techniques of DNA isolation and identification of three Streptococci bacteria by Polimerase Chain Reaction (PCR) was made. Results: We identified Streptococcus salivarius and Streptococcus gordoni only in the lower interincisal region. These species were also present mainly in the first permanent non-decayed molars. On the contrary, Streptococcus mutans was found mostly in decayed first permanent molars. Conclusion: This preliminary study establishes a possible association between the absence of dental caries and the presence of Streptococcus gordonii and Streptococcus salivarius. Since these two species are described as alkali producers, it is suggested that their presence somehow confers protection against caries. These results support new dental caries prevention strategies based on oral biofilm modulation by enrichment with alkalinogenic species.

Keywords: dental caries, oral biofilm, Streptococcus gordonii, Streptococcus salivarius

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Authors: Aqeela Ashraf, Muhammad Imran, Tahir Yaqub, Muhammad Tayyab, Yung Fu Chang

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For the identification of bovine mastitic pathogen, an economical, rapid and sensitive molecular diagnostic assay is developed by PCR multiplexing of gene and pathogenic species specific DNA sequences. The multiplex PCR assay is developed for detecting nine important bacterial pathogens causing mastitis Worldwide. The bacterial species selected for this study are Streptococcus agalactiae, Streptococcus dysagalactiae, Streptococcus uberis, Staphylococcus aureus, Escherichia coli, Staphylococcus haemolyticus, Staphylococcus chromogenes Mycoplasma bovis and Staphylococcus epidermidis. A single reaction assay was developed and validated by 27 reference strains and further tested on 276 bacterial strains obtained from culturing mastitic milk. The multiplex PCR assay developed here is further evaluated by applying directly on genomic DNA isolated from 200 mastitic milk samples. It is compared with bacterial culturing method and proved to be more sensitive, rapid, economical and can specifically identify 9 bacterial pathogens in a single reaction. It has detected the pathogens in few culture negative mastitic samples. Recognition of disease is the foundation of disease control and prevention. This assay can be very helpful for maintaining the udder health and milk monitoring.

Keywords: multiplex PCR, bacteria, mastitis, milk

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Authors: Ayman El Behiry, Rasha Nabil Zahran, Reda Tarabees, Eman Marzouk, Musaad Al-Dubaib

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Mastitis is one of the most economic disease affecting dairy cows worldwide. Its classic diagnosis using bacterial culture and biochemical findings is a difficult and prolonged method. In this research, using of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) permitted identification of different microorganisms with high accuracy and rapidity (only 24 hours for microbial growth and analysis). During the application of MALDI-TOF MS, one hundred twenty strains of Staphylococcus and Streptococcus species isolated from milk of cows affected by clinical and subclinical mastitis were identified, and the results were compared with those obtained by traditional methods as API and VITEK 2 Systems. 37 of totality 39 strains (~95%) of Staphylococcus aureus (S. aureus) were exactly detected by MALDI TOF MS and then confirmed by a nuc-based PCR technique, whereas accurate identification was observed in 100% (50 isolates) of the coagulase negative staphylococci (CNS) and Streptococcus agalactiae (31 isolates). In brief, our results demonstrated that MALDI-TOF MS is a fast and truthful technique which has the capability to replace conventional identification of several bacterial strains usually isolated in clinical laboratories of microbiology.

Keywords: identification, mastitis pathogens, mass spectral, phenotypical

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Authors: Aliya Tokbergenova, Maida Tusupbekova, Daulet Dzhangaliyev, Alena Lavrinenko

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Background: Odontogenic phlegmons are ranked the first among pyoinflammatory processes in the frequency of hospitalization in maxillofacial surgery in the post-Soviet countries. The main role in etiology is played by obligate anaerobes and aerobes. According to numerous data, the structure of aerobic pathogens is dominated by staphylococci and gram-negative bacteria. Aim: The research aim is to study the microflora of the purulent discharge odontogenic inflammatory processes. Materials and methods: A total of 220 patients have been examined, of which 120 patients aged 25-59 years have been included in the research who did not have comorbidity hospitalized in the maxillofacial hospital in Karaganda (Kazakhstan) from January 2016 to July 2017. The bacteriological research has been carried out on the basis of the multiaccess laboratory of the KSMU, through the Matrix Assisted Laser Desorption/Ionization (MALDI) apparatus. The material sample was pus from the inflammation focus, taken during the operating period. Results: According to the research among 120 patients (100%), 15 patients (12.5%) have had microorganisms not grown. From 105 (87.5%) bacteriological results, it has been revealed the following 1) Streptococcus: 51 (42.5%): Streptococcus beta-haemolytic: 17 (14.2%), Streptococcus pneumoniae: 12 (10%), Streptococcus anginosus: 8 (6.6%), Streptococcus oralis: 8 (6.6%), Streptococcus constellatus: 6 (5.0%); 2) Staphylococci: 27 (22.5%): Staphylococci aureus: 14 (11.7%) and Staphylococci epidermidis: 13 (10.8%); 3) Pseudomonas aeruginosa: 12 (10%); 4) Neisseria: 11 (9.1%): Neisseria mucosa: 5 (4.1%) and Neisseria macacae: 6 (5.0%); 5) Klebsiella pneumoniae: 2 (1.7%); 6) Stenotrophomonas maltophilia: 2 (1.7%). 15 patients (12.5%) experienced complications in the form of 1) The dissemination of the process in 10 patients (8.4%). 2) Osteomyelitis in 3 (2.5%). 3) Mediastinitis in 1 (0.8%). 4) Sinusitis in 1 (0.8%). 15 patients (100%) were carried out repeated bacteriological examination, the following was revealed: 1) Streptococcus: 10 (66.7%): Streptococcus beta-haemolytic: 4 (26.7%), Streptococcus pneumoniae: 2 (13.3%), Streptococcus аnginosus: 2 (13.3%), Streptococcus oralis: 1 (6.7%), Streptococcus constellatus: 1 (6.7%); 2) Staphylococci: 4 (26.7%): Staphylococci aureus: 3 (20%) and Staphylococci epidermidis: 1 (6.7%); 3) Pseudomonas aeruginosa: 1 (6.7%). Conclusions: Thus, according to our research data, streptococci predominate in the odontogenic processes microflora in aerobic flora in the central Kazakhstan region, which refutes the leading role of staphylococci in the development of odontogenic inflammatory processes, thus creating prerequisites for studying new treatment approaches.

Keywords: maxillofacial surgery, microflora, odontogenic phlegmons, pyo-inflammatory

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Authors: Nenny Harijani, Dhandy Koesoemo Wardhana

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The aim of this study is to know the bacteriocin as antimicrobial activity produced by Lactic Acid Bacteria (LAB) as Antibacterial of Sub Clinic Mastitis on Dairy Cows. The antimicrobial is produced by LAB which isolates from cattle intestine can inhibit the growth Staphylococcus aureus, Steptocococcus agalactiae an Escherichia coli which were caused by dairy cattle subclinical mastitis. The failure of this bacteria growth was indicated by the formation of a clear zone surrounding the colonies on Brain Heart Infusion Agar plate. The bacteriocin was produced by Lactic Acid Bacteria (LAB) as antimicrobial, which could inhibit the growth of indicator bacteria Staphylococcus aureus, S.aglactiae and E.coli. This study was also developed bacteriocin to be used as a therapeutic of subclinical mastitis on dairy cows. The method used in this study was isolation, selection and identification of LAB using Mann Rogosa Sharp Medium, followed by characterization of the bacteriocin produced by LAB. The result of the study showed that bacteriocin isolated from beef cattle’s intestine could inhibit the growth Staphylococcus aureus, S. agalactiae, an Escherichia coli, which was indicated by clear zone surrounding the colonies on Brain Heart Infusion Agar plate. Characteristics of bacteriocin were heat-stable exposed to 80 0C for 30 minutes and 100 ⁰C for 15 minutes and inactivated by proteolytic enzymes such as trypsin. This approach has suggested the development of bacteriocin as a therapeutic agent for subclinical mastitis in dairy cattle.

Keywords: lactic acid bacteria, bacteriocin, staphylococcus aureus, S. agalactiae, E. coli, sub

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Authors: Djeddi Khaled, Houssou Hind, Miloudi Abdelatif, Rabah Siham

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Camels play a vital role as multipurpose animals, providing milk meat and serving as a means of transportation. They serve as a financial reserve for pastoralists and hold significant cultural and social value. Camel milk, known for its exceptional nutritional properties, is considered a valuable substitute for human milk. However, udder infections, particularly mastitis, pose significant challenges to camel farming. Clinical and subclinical mastitis can lead to substantial economic losses. Mastitis, especially the subclinical form, is a persistent and prevalent condition affecting milk hygiene and quality in dairy camels. This review offers insights into the prevalence and risk factors associated with subclinical mastitis in camels. The prevalence of subclinical mastitis in dairy camels was found to range from 9.28% to 87.78%. Major pathogens responsible for camel mastitis include Staphylococcus aureus, Coagulase-negative Staphylococcus, Streptococcus agalactiae, Streptococcus dysgalactiae, Escherichia coli, Micrococcus spp, Pasteurella haemolytica and Corynebacterium spp. The study outlines key risk factors contributing to camel mastitis, emphasizing factors such as severe tick infestation, age, stage of lactation, parity, body condition score, skin lesion on the teats or udders, anti-suckling devices, previous history of the udder, conformation of the udder, breed, unhygienic milking practices, production system, amongst others have been reported to be important in the prevalence of subclinical mastitis. This comprehensive overview provides valuable insights into the multifaceted aspects of camel mastitis, encompassing prevalent bacterial pathogens and diverse risk factors. The findings underscore the importance of holistic management practices, emphasizing hygiene, health monitoring, and targeted interventions to ensure the well-being and productivity of camels in various agro-pastoral contexts.

Keywords: bacterial pathogens, camel, mastitis, risk factors

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Authors: Abhichet Nobhiwong, Jiraporn Rojtinnakorn, Udomluk Sompong

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Six probiotic strains isolated from Chiang Mai and Chiang Rai province, Thailand; CR1-2, CM3-4, CM5-2, CR7-8, CM10-5 and CM10-8 were used to study their morphology and inhibition activity on three pathogenic bacteria; Aeromonas sp., Streptococcus sp. and Flavobacterium sp. that isolated from infected Nile tilapia. The agar well diffusion technique was applied for 24 and 48 hours incubation. Interestingly, some probiotics showed good inhibition activity both 24 and 48 hours on each 3 bacterial pathogens. The capable inhibiting Aeromonas sp. were CR1-2 and CR5-2 with inhibition diameters of 13.0 mm and 11.2 mm, respectively. For Streptococcus sp., effective probiotics were CR10-2 with inhibition diameters of 10.7 mm. Whereas for Flavobacterium sp., effective probiotics were CR5-2 with inhibition diameter of 9.7 mm. It can be concluded that these probiotics have potentiality to develop as the pathogens biocontrol products. These will be support for safety and organic aquaculture that which the most worthy for people health.

Keywords: probiotics, Aeromanas sp., Streptococcus sp., Flavobacterium sp.

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Authors: Chougrani Fadela, Cheriguene Abderrahim

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Fifty three strains of thermophilic and mesophilic lactic acid bacteria were isolated from the ewe’s milk. Identification reveals the presence of nineteen strains (36%) of Lactobacillus sp., seventeen strains (32%) of Lactococcus sp., nine strains (17%) of Streptococcus thermophilus and eight strains (15%) of Leuconostoc sp. The strains were characterized for their technological properties. A high diversity of properties among the studied strains was demonstrated. On the basis of technological characteristics, two strains (Lactobacillus bulgaricus and Streptococcus thermophilus) were screened with respect to their acid and flavour production for the preparation of a natural yogurt and compared to a commercial starter cultures. Sensorial analyses revealed that the product manufactured on the basis of the isolated strains have a cohesiveness and adhesiveness corresponding to standard products. The pH and the acidity recorded are also within accepted levels during all the period of conservation.

Keywords: Lactobacillus bulgaricus, Streptococcus thermophilus, yoghurt, cohesiveness, adhesiveness, Algerian ewe’s milk

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Authors: Negar Zaheddoust, Negin Zaheddoust, Abbas Asoudeh-Fard

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Probiotic bacteria have been employed as a novel and less side-effect strategy for anticancer therapy. Since the oral cavity is a host for probiotic and pathogen bacteria to colonize, more investigation is needed to evaluate the effectiveness of this novel adjunctive treatment for oral cancer. We considered Lactobacillus plantarum as a probiotic and Streptococcus mutans as a pathogen bacterium in our study. The aim of this study is to examine the effect of Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT / PTEN, and MAPK signaling pathway, which is involved in apoptosis or survival of oral cancer KB cells. On the other hand, to study the effects of these bacteria on normal cells, we used HUVEC cells. The KB and HUVEC cell lines were co-cultured with Lactobacillus plantarum and Streptococcus mutans isolated from traditional Iranian dairy and dental plaque, respectively. The growth-inhibitory effects of these two bacteria on KB and HUVEC cells were determined by (3-(4, 5-dimethylthiazolyl-2)-2,5diphenyltetrazolium bromide) MTT assay. MTT results demonstrated that the proliferation of KB cells was affected in a time, dose, and strain-dependent manner. In the following, the examination of induced apoptosis or necrosis in co-cultured KB cells with the best IC50 concentration of the Lactobacillus plantarum and Streptococcus mutans will be analyzed by FACS flow cytometry, and the changes in gene expression of Casp3, AKT / PTEN, MAPK genes will be evaluated using real-time polymerase chain reaction.

Keywords: cancer therapy, induced apoptosis, oral cancer, probiotics

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Authors: Adila Shaukat, Hussam Al Soub, Muna Al Maslamani, Abdullatif Al Khal

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Background: The aim of this study was to assess clinical presentation and antimicrobial susceptibility of Streptococcus (S.) anginosus group infections in Hamad General Hospital, a tertiary care hospital in the state of Qatar, which is a multinational community. The S. anginosus group is a subgroup of viridans streptococci that consist of 3 different species: S. anginosus, S. constellatus, and S. intermedius. Although a part of the human bacteria flora, they have potential to cause suppurative infections. Method: We studied a total of 101 patients with S. anginosus group infections from January 2006 until March 2012 by reviewing medical records and identification of organisms by VITEK 2 and MALDI-TOF. Results: The most common sites of infection were skin and soft tissue, intra-abdominal, and bacteremia (28.7%, 24.8%, and 22.7%, respectively). Abscess formation was seen in approximately 30% of patients. Streptococcus constellatus was the most common isolated species (40%) followed by S. anginosus(30%) and S. intermedius(7%). In 23% of specimens, the species was unidentified. The most common type of specimen for organism isolation was blood followed by pus and tissue (50%, 22%, and 8%, respectively). Streptococcus constellatus was more frequently associated with abdominal and skin and soft tissue infections than the other 2 species, whereas S. anginosus was isolated more frequently from blood. All isolates were susceptible to penicillin, ceftriaxone, and vancomycin. Susceptibility to erythromycin and clindamycin was also good, reaching 91% and 95%, respectively. Forty percent of patients needed surgical drainage along with antibiotic therapy. Conclusions: Identification of S. anginosus group to species level is helpful in clinical practice because different species exhibit different pathogenic potentials.

Keywords: abscess, bacterial infection, bacteremia, Streptococcus anginosus

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Authors: Ahmed Jibrin Uttu, Maimuna Waziri

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The root bark of Glossonema boveanum (Decne), a member of Apocynaceae family, is used by traditional medicine practitioner to treat urinary and respiratory tract infections, bacteremia, typhoid fever, bacillary dysentery, diarrhea and stomach pain. This present study aims to validate the medicinal claims ascribed to the root bark of the plant. Preliminary phytochemical study of the root bark extracts (n-hexane, ethyl acetate, chloroform and methanol extracts) showed the presence of alkaloids, carbohydrates, steroids, triterpenes, cardiac glycosides, saponins, tannins and flavonoids. Antimicrobial study of the extracts showed activities against Staphylococus aureus, Bacillus subtilis, Salmonella typhii, Shigella dysenteriae, Escherichia coli, Enterobacter cloacae, Streptococcus agalactiae and Candida albicans while Micrococcus luteus, Pseudomonas aeruginosa and Klebsiella Pneumoniae showed resistance to all the extracts. The inhibitory effect was compared with the standard drug ciprofloxacin and fluconazole. MIC and MBC for both extracts were also determined using the tube dilution method. This study concluded that the root bark of G. boveanum, used traditionally as a medicinal plant, has antimicrobial activities against some causative organisms.

Keywords: Glossonema boveanum (Decne.), phytochemical, antimicrobial, minimum inhibitory concentration, minimum bactericidal concentration

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Authors: Kanchana Sitlaothaworn

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Yogurt capsule was made by mixing 14% w/v of reconstitution of skim milk with 2% FOS. The mixture was fermented by commercial yogurt starter comprising Lactobacillus bulgaricus and Streptococcus thermophilus. These yogurts were made as yogurt powder by freeze-dried. Yogurt powder was put into capsule then stored for 28 days at 4oc. 8ml of commercial yogurt was found to be the most suitable inoculum size in yogurt production. After freeze-dried, the viability of L. bulgaricus and S. thermophilus reduced from 109 to 107 cfu/g. The precence of sucrose cannot help to protect cell from ice crystal formation in freeze-dried process, high (20%) sucrose reduced L. bulgaricus and S. thermophilus growth during fermentation of yogurt. The addition of FOS had reduced slowly the viability of both L. bulgaricus and S. thermophilus similar to control (without FOS) during 28 days of capsule storage. The viable cell exhibited satisfactory viability level in capsule storage (6.7x106cfu/g) during 21 days at 4oC.

Keywords: yogurt capsule, Lactobacillus bulgaricus, Streptococcus thermophilus, freeze-drying, sucrose

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Authors: Agampodi Promoda Perera, Yong Shin, Mi Kyoung Park

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The successful detection of pathogenic bacteria in blood provides important information for early detection, diagnosis and the prevention and treatment of infectious diseases. Silicon microring resonators are refractive-index-based optical biosensors that provide highly sensitive, label-free, real-time multiplexed detection of biomolecules. We demonstrate the technique of using GO (graphene oxide) to enhance the signal output of the silicon microring optical sensor. The activated carboxylic groups in GO molecules bind directly to single stranded DNA with an amino modified 5’ end. This conjugation amplifies the shift in resonant wavelength in a real-time manner. We designed a capture probe for strain Staphylococcus aureus of 21 bp and a longer complementary target sequence of 70 bp. The mismatched target sequence we used was of Streptococcus agalactiae of 70 bp. GO is added after the complementary binding of the probe and target. GO conjugates to the unbound single stranded segment of the target and increase the wavelength shift on the silicon microring resonator. Furthermore, our results show that GO could successfully differentiate between the mismatched DNA sequences from the complementary DNA sequence. Therefore, the proposed concept could effectively enhance sensitivity of pathogen detection sensors.

Keywords: label free biosensor, pathogenic bacteria, graphene oxide, diagnosis

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Authors: Mohd Adil, Rosina Khan, Asad U. Khan, Vasantha Rupasinghe HP

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The study examined the effects of Syzygium aromaticum extracts on the virulence properties of Streptococcus mutans. The activity of glucosyltransferases in the presence of crude and diethylether fraction was reduced to 80% at concentration 78.12μg/ml and 39.06μg/ml respectively. The glycolytic pH drop by S. mutans cells was also disrupted by these extracts without affecting the bacterial viability. Microscopic analysis revealed morphological changes of the S. mutans biofilms, indicating that these plant extracts at sub-MICs could significantly affect the ability of S. mutans to form biofilm with distorted extracellular matrix. Furthermore, with the help of quantitative RT-PCR, the expression of different genes involved in adherence, quorum sensing, in the presence of these extracts were down regulated. The crude and active fractions were found effective in preventing caries development in rats. The data showed that S. aromaticum holds promise as a naturally occurring source of compounds that may prevent biofilm-related oral diseases.

Keywords: biofilm, quorum sensing, Streptococcus mutans, Syzygium aromaticum extract

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Authors: M. H. Nahaisi, N. M. Almaroum

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Laban is a Libyan traditional fermented milk product. This lactic fermentation has been known in many cities of Libya long time ago as stable, nutritious, refreshing drink especially during the summer. 16 naturally fermented milk samples were collected from different cities located in North West of Libya. The average pH, titratable acidity, fat and total solids were 4.16, 0.73%, 1.54% and 8.12 % respectively. Coliform, yeast and mold counts were 21×10⁴, 39×10⁴ and 41 ×10³ cfu/ ml. respectively. The average Lactococcus, Streptococcus, Mesophilic Lactobacillus / Leuconostoc and Thermophilic Lactobacillus counts were 99 ×10⁷, 96 ×10⁷, 93 ×10⁷ and 15 ×10⁷ cfu / ml. respectively. A total of one hundred forty two lactic acid bacteria (LAB) isolates were identified to the genus level as Lactobacillus (48.59%), Lactococcus (43.66%), Streptococcus (4.93%) and Leuconostoc (2.82%). Sugar fermentation tests have revealed that the most frequently Lactobacillus species was found to be Lactobacillus delbrueckii ssp. lactis (62.32%) followed by Lactobacillus plantarum (31.88%). Furthermore, other selected LAB isolates were identified by API 50 CH test as Lactococcus lactis ssp. lactics, Lactobacillus pentosus, Lactobacillus brevis and Leuconostoc mesenteroides ssp. cremoris.

Keywords: traditional fermented milk, laban, lactococcus, streptococcus, mesophilic lactobacillus, thermophilic lactobacillus counts

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Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii

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A huge revolution has emerged in genome engineering by the discovery of CRISPR (clustered regularly interspaced palindromic repeats) and CRISPR-associated system genes (Cas) in bacteria. The function of type II Streptococcus pyogenes (Sp) CRISPR/Cas9 system has been confirmed in various species. Other S. thermophilus (St) CRISPR-Cas systems, CRISPR1-Cas and CRISPR3-Cas, have been also reported for preventing phage infection. The CRISPR1-Cas system interferes by cleaving foreign dsDNA entering the cell in a length-specific and orientation-dependant manner. The S. thermophilus CRISPR3-Cas system also acts by cleaving phage dsDNA genomes at the same specific position inside the targeted protospacer as observed in the CRISPR1-Cas system. It is worth mentioning, for the effective DNA cleavage activity, RNA-guided Cas9 orthologs require their own specific PAM (protospacer adjacent motif) sequences. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of target site for the three orthogonals of Cas9 protein, a well-organized procedure will be required for high-throughput and accurate mining of possible target sites in a large genomic dataset. Consequently, we created a reliable procedure to explore potential gRNA sequences for type I (Streptococcus thermophiles), II (Streptococcus pyogenes), and III (Streptococcus thermophiles) CRISPR/Cas systems. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows: i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. The output of such procedure highlights the power of comparative genome mining for different CRISPR/Cas systems. This could yield a repertoire of Cas9 variants with expanded capabilities of gRNA design, and will pave the way for further advance genome and epigenome engineering.

Keywords: CRISPR/Cas systems, gRNA mining, Streptococcus pyogenes, Streptococcus thermophiles

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Authors: Marta Skwarecka, Patrycja Bloch, Rafal Walkusz, Oliwia Urbanowicz, Grzegorz Zielinski, Sabina Zoledowska, Dawid Nidzworski

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Upper respiratory tract infections are one of the most common reasons for visiting a general doctor. Streptococci are the most common bacterial etiological factors in these infections. There are many different types of Streptococci and infections vary in severity from mild throat infections to pneumonia. For example, S. pyogenes mainly contributes to acute pharyngitis, palatine tonsils and scarlet fever, whereas S. Streptococcus pneumoniae is responsible for several invasive diseases like sepsis, meningitis or pneumonia with high mortality and dangerous complications. There are only a few diagnostic tests designed for detection Streptococci from the infected throat of patients. However, they are mostly based on lateral flow techniques, and they are not used as a standard due to their low sensitivity. The diagnostic standard is to culture patients throat swab on semi selective media in order to multiply pure etiological agent of infection and subsequently to perform antibiogram, which takes several days from the patients visit in the clinic. Therefore, the aim of our studies is to develop and implement to the market a Point of Care device for the rapid identification of Streptococcus pyogenes and Streptococcus pneumoniae with simultaneous identification of antibiotic resistance genes. In the course of our research, we successfully selected genes for to-species identification of Streptococci and genes encoding antibiotic resistance proteins. We have developed a reaction to amplify these genes, which allows detecting the presence of S. pyogenes or S. pneumoniae followed by testing their resistance to erythromycin, chloramphenicol and tetracycline. What is more, the detection of β-lactamase-encoding genes that could protect Streptococci against antibiotics from the ampicillin group, which are widely used in the treatment of this type of infection is also developed. The test is carried out directly from the patients' swab, and the results are available after 20 to 30 minutes after sample subjection, which could be performed during the medical visit.

Keywords: antibiotic resistance, Streptococci, respiratory infections, diagnostic test

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Authors: Shiva Emami, Jenny Persson, Bengt Johansson Lindbom

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Group A streptococcus (GAS) is a prevalent human pathogen, causing a wide range of infections and diseases. One of the most well-known virulence factors in GAS is M protein, a surface protein that facilitates bacterial invasion. In this study, we used a recombinant GAS strain (GAS-2W) expressing M protein containing a hyper immunogenic peptide (2W). Mice were immunized three times with heat-killed-GAS subcutaneously at three weeks intervals. Three weeks post last immunization, mice were challenged intraperitoneally with a lethal dose of live GAS. In order to investigate the impact of IFN-ƴ and antibodies in protection against GAS infection, we used a mouse model knock-out for IFN-ƴ (IFN-ƴ KO). We observed immunization with GAS-2W strain can increase protection against GAS infection in mice compared with the original GAS strain. Higher levels of antibodies against M1 protein were measured in GAS-2W-immunized mice. There was also a significant increase in IgG2c response in mice immunized with GAS2W. By using IFN-ƴ KO mice, we showed that not a high level of total IgG, but IgG2c was correlated with protection through the i.p challenge. It also emphasizes the importance of IFN-ƴ cytokine to combat GAS by isotype switching to IgG2c (which is opsonic for phagocytosis). Our data indicate the crucial role of IFN-ƴ in the protective immune response that, together with IgG2c, can induce protection against GAS.

Keywords: Group A streptococcus, IgG2c, IFN-γ, protection

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Authors: Albert Chang, Duncan Webster

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Streptococcus equi subspecies zooepidemicus infections in humans can be contracted through contact with domestic animals or unpasteurized dairy products. Although infection in humans is rare, the course can be fulminant. We describe the case of a 75-year-old, immunocompetent male, who developed disseminated disease with bacteremia, native aortic valve endocarditis, suppurative pericarditis with cardiac tamponade, meningitis and bilateral endopthalmitis. Despite treatment with pericardial drain placement, intravenous ceftriaxone and rifampin the patient, unfortunately, did not survive. To date, reported cases of disseminated infection by S. zooepidemicus are few. Furthermore, with the review of the literature, this case demonstrates the broadest organ system involvement reported. Of interest, previous studies have suggested an affinity of this organism for certain organ systems and this case corroborates an emerging association of S. zooepidemicus with endopthalmitis. In addition, this is the second Canadian case of documented human infection with both cases being similar in clinical features, presentation, and geographical location. A discussion regarding previous S. zooepidemicus outbreaks and the potential for zoonotic outbreaks to occur is included. In short, this case report should serve to warn clinicians regarding complications and sites of haematogenous seeding in the setting of disseminated S. zooepidemicus infections.

Keywords: endopthalmitis, endocarditis, meningitis, Streptococcus equi subspecies zooepidemicus

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Authors: Martynova Alina, Lyubov Buzoleva

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Streptococcus pneumoniae is the causative agent of pneumonias and meningitids throught all the world. Having high genetic diversity, this microorganism can cause different clinical forms of pneumococcal infections and microbiologically it is really difficult diagnosed by routine methods. Also, epidemiological surveillance requires more developed methods of molecular typing because the recent method of serotyping doesn't allow to distinguish invasive and non-invasive isolates properly. Non-coding genome of bacteria seems to be the interesting source for seeking of highly distinguishable markers to discriminate the subspecies of such a variable bacteria as Streptococcus pneumoniae. Technically, we proposed scheme of discrimination of S.pneumoniae strains with amplification of non-coding region (SP_1932) with the following restriction with 2 types of enzymes of Alu1 and Mn1. Aim: This research aimed to compare different methods of typing and their application for molecular epidemiology purposes. Methods: we analyzed population of 100 strains of S.pneumoniae isolated from different patients by different molecular epidemiology methods such as pulse-field gel electophoresis (PFGE), restriction polymorphism analysis (RFLP) and multilolocus sequence typing (MLST), and all of them were compared with classic typing method as serotyping. The discriminative power was estimated with Simpson Index (SI). Results: We revealed that the most discriminative typing method is RFLP (SI=0,97, there were distinguished 42 genotypes).PFGE was slightly less discriminative (SI=0,95, we identified 35 genotypes). MLST is still the best reference method (SI=1.0). Classic method of serotyping showed quite weak discriminative power (SI=0,93, 24 genotypes). In addition, sensivity of RFLP was 100%, specificity was 97,09%. Conclusion: the most appropriate method for routine epidemiology surveillance is RFLP with non-coding region of Streptococcsu pneumoniae, then PFGE, though in some cases these results should be obligatory confirmed by MLST.

Keywords: molecular epidemiology typing, non-coding genome, Streptococcus pneumoniae, MLST

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Authors: Nadia A. Mohamed, Magdy W. Sabaa, Ahmed H. H. El-Ghandour, Marwa M. Abdel-Aziz, Omayma F. Abdel-Gawad

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Eighteen carboxymethyl chitosan (CMCh) schiff bases and their reduced derivatives have been synthesized. They were characterized by spectral analyses (FT-IR and H1-NMR) and scanning electron microscopy observation. Their antibacterial activities against Streptococcus pneumoniae (RCMB 010010), Bacillis subtilis (RCMB 010067), as Gram positive bacteria and Escherichia coli (RCMB 010052) as Gram negative bacteria and the antifungal activity against Aspergillus fumigatus (RCMB 02568), Geotricum candidum (RCMB 05097), and Candida albicans (RCMB 05031) were examined using agar disk diffusion method. The results demonstrate how the antibacterial and the antifungal activity are clearly affected by both the nature and position of the substituent groups in the aryl ring of the prepared derivatives. CMCh-4-nitroBenz Schiff base and its reduced form show higher antimicrobial activity comparing with other para substituted derivatives. CMCh-4-nitroBenz Schiff base: 18.3, 17, and 15.6 mm against Bacillis subtilis, Streptococcus pneumonia, and Escherichia coli respectively and 16.2, 17.3, and 16.4 mm against Aspergillus fumigates, Geotricum candidum, and Candida albicans respectively. CMCh-4-nitroBenz reduced form: 19.5, 18.7, and 16.2 mm against Bacillis subtilis, Streptococcus pneumonia, and Escherichia coli respectively and 17.5, 19.5, and 17.4 mm against Aspergillus fumigates, Geotricum candidum, and Candida albicans respectively. Also CMCh-3-bromoBenz show good results; CMCh-3-bromoBenz schiff base: 19.2, 16.9, and 14.6 mm Bacillis subtilis, Streptococcus pneumonia, and Escherichia coli respectively and 18.4, 17.6, and 15.9 mm against Aspergillus fumigates, Geotricum candidum, and Candida albicans respectively.

Keywords: chitosan, schiff base, minimum inhibition concentration, antimicrobial activity

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Authors: Amalia Dwi Berliyanti Amel

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Streptococcus mutants bacteria are bacteria that are believed to be the cause of the growth of dental plaque which can further adversely affect dental caries if left unchecked. Previous research has shown that Moringa leaf extract can slow down the growth rate of this bacterium. This study aims to make the best formulation of mouthwash with the active ingredient of Moringa leaf extract based on its antibacterial and organoleptic test results. Nine mouthwash variations were carried out with two factors and three levels, namely a comparison of the concentration of sorbitol (A) with three levels namely 15% (A1), 20% (A2), and 25% (A3), and peppermint added (B) with three levels, namely 0.2% (B1), 0.25% (B2), and 0.3% (B3). The test parameters performed as the determination of the best mouthwash are based on physicochemical properties which include pH and viscosity as well as organoleptic test results which include color, viscosity, aroma, taste, sensation in the mouth, and general appearance. The results showed that the bright zone as a test for the antibacterial activity of Streptococcus mutants began to be seen at a concentration of 5%. Moringa leaf mouthwash formulation has a pH value between 6 - 7, with a control of 6. Whereas the mucosa leaf mouthwash vascularity produced between 1.1 - 1.7 cP with a control of 1.1 cP. Moringa leaf mouthwash and control have the same total number of microbes, namely 0 colonies / mL. Based on organoleptic tests performed with 20 panelists, it was shown that the best mouthwash formulation was formulation A1B3 with sorbitol composition 15% and peppermint 0.3%.

Keywords: antibasteria, formula, moringa leaf, mouthwash

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Authors: A. T. Laiche, M. L. Tlil, Benine B., S. Bechoua

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The aim of this study is to isolate and select, from camel milk from El-Oued region in Algeria, a strains of lactic acid bacteria and possessing probiotic properties ; and to evaluate their potential effect on intestinal disorders in Wistar ratsmThe results relating to the selection of probiotic strains confirms that the Thermophilic streptococcus exhibits the best probiotic activity performance, with a resistance important to different degrees of pH and to bile salts, and a remarkable antibacterial activity and resistance to antibiotics compared to the other four isolated strains. In the in vivo study, diseases are induced in rats at the level of the digestive system, it was reported that the administration of Escherichia coli and castor oil caused an intestinal disorders. The microscopic observation of the histological section of the intestine showed a damaged intestinal structure and some symptoms of its irritation, including a decrease in the height of the villi and the presence of others destroyed cells, and after treatment with Streptococcus thermophilus, the microscopic observation of the cut of the histological section of the intestine showed almost complete disappearance of the mentioned symptoms, The dosage of the hematological parameters by complete blood count (CBC) is in agreement with the results of the histological sections.

Keywords: camel milk, probiotic, pathogenic bacteria, intestinal disorders, lactic acid bacteria

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Authors: Andreev V. A., Kovalenko T. N., Privolnev V. V., Chernavin A. V., Knyazeva E. R.

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Aim: To evaluate the antagonistic activity of Streptococcus salivarius K12 (SsK12) against ENT and oral cavity infection pathogens (S. pneumoniae, S. pyogenes, S. aureus), gram-negative bacteria (E. coli, P. aeruginosa) and C. albicans. Materials and methods: The probiotic strain SsK12 was isolated from the dietary supplement containing at least 1 × 109 CFU per tablet. The tablet was dissolved in the enrichment broth. The resulting suspension was seeded on 5% blood agar and incubated at 35°C in 4-6% CO2 for 48 hours. The raised culture was identified as Streptococcus salivarius with MALDI-TOF mass spectrometry method. The evaluation of SsK12 antagonistic activity was carried out using a perpendicular streak technique. The daily SsK12 culture was inoculated as heavy streaks with a loop at one side of Petri dish with the Muller-Hinton agar (MHA) and incubated for 24 hours at 350 C in anaerobic conditions. It was supposed that bacteriocins would diffuse over the whole area of the agar media. On the next day S. pneumoniae, S. pyogenes, S. aureus, E. coli, P. aeruginosa and C. albicans clinical isolates were streaked at the clear side of MHA Petri dish. MHA Petri dish inoculated with SsK12 (one part) and with the respective clinical isolates (another part) streaked perpendicularly on the same day was used as the control. Results: There was no growth of S. pyogenes on the Petri dish with SsK12 daily culture; the growth of a few colonies of S. pneumonia was noted. The growth of S. aureus, E. coli, P. aeruginosa and C. albicans was noted along the inoculated streak. On the control Petri dish with simultaneous inoculating of the SsK12 strain and the test cultures, the growth of all the testes isolates was noted. Conclusions: (1) SsK12 possesses perfect antagonistic activity against S. pyogenes and good activity against S. pneumoniae. (2) There was no antagonistic activity of SsK12 against S. aureus, E. coli, P. aeruginosa and C. albicans. (3) SsK12 antagonistic properties make it possible to use this probiotic strain for prophylaxis of recurrent ENT infections.

Keywords: probiotics, SsK12, streptococcus salivarius K12, antagonistic activity

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Authors: Soledad Carinelli, Iñigo Fernández, José Luis González-Mora, Pedro A. Salazar-Carballo

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Mastitis is the most relevant inflammatory disease in cattle, affecting the animal health and causing important economic losses on dairy farms. This disease takes place in the mammary gland or udder when some opportunistic microorganisms, such as Staphylococcus aureus, Streptococcus agalactiae, Corynebacterium bovis, etc., invade the teat canal. According to the severity of the inflammation, mastitis can be classified as sub-clinical, clinical and chronic. Standard methods for mastitis detection include counts of somatic cells, cell culture, electrical conductivity of the milk, and California test (evaluation of “gel-like” matrix consistency after cell lysed with detergents). However, these assays present some limitations for accurate detection of subclinical mastitis. Currently, haptoglobin, an acute phase protein, has been proposed as novel and effective biomarker for mastitis detection. In this work, an electrochemical biosensor based on polydopamine-modified magnetic nanoparticles (MNPs@pDA) for haptoglobin detection is reported. Thus, MNPs@pDA has been synthesized by our group and functionalized with hemoglobin due to its high affinity to haptoglobin protein. The protein was labeled with specific antibodies modified with alkaline phosphatase enzyme for its electrochemical detection using an electroactive substrate (1-naphthyl phosphate) by differential pulse voltammetry. After the optimization of assay parameters, the haptoglobin determination was evaluated in milk. The strategy presented in this work shows a wide range of detection, achieving a limit of detection of 43 ng/mL. The accuracy of the strategy was determined by recovery assays, being of 84 and 94.5% for two Hp levels around the cut off value. Milk real samples were tested and the prediction capacity of the electrochemical biosensor was compared with a Haptoglobin commercial ELISA kit. The performance of the assay has demonstrated this strategy is an excellent and real alternative as screen method for sub-clinical bovine mastitis detection.

Keywords: bovine mastitis, haptoglobin, electrochemistry, magnetic nanoparticles, polydopamine

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Authors: Hamza A. Pantami, Khozizah Shaari, Intan S. Ismail, Chong C. Min

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Introduction: Tilapia is the second-highest harvested freshwater fish species in Malaysia, available in almost all fish farms and markets. Unfortunately, tilapia culture in Malaysia is highly affected by Aeromonas hydrophila and Streptococcus agalactiae, which affect the production rate and consequently pose a direct negative economic impact. Reliance on drugs to control or reduce bacterial infections has been led to contamination of water bodies and development of drug resistance, as well as gave rise to toxicity issues in downstream fish products. Resorting to vaccines have helped curb the problem to a certain extent, but a more effective solution is still required. Using microalgae-based feed to enhance the fish immunity against bacterial infection offers a promising alternative. Objectives: This study aims to evaluate the efficacy of Chlorella vulgaris at lower percentage incorporation in feeds for an immune boost of tilapia in a shorter time. Methods: The study was in two phases. The safety concentration studies at 500 mg/kg-1 and the administration of cultured C. vulgaris biomass via incorporation into fish feed for five different groups in three weeks. Group 1 was the control (0% incorporation), whereas group 2, 3, 4 and 5 received 0.625%, 1.25%, 2.5% and 5% incorporation respectively. The parameters evaluated were the blood profile, serum lysozyme activity (SLA), serum bactericidal activity (SBA), phagocytosis activity (PA), respiratory burst activity (RBA), and lymphoproliferation activity (LPA). The data were analyzed via ANOVA using SPSS (version 16). Further testing was done using Tukey’s test. All tests were performed at the 95% confidence interval (p < 0.05). Results: There were no toxic signs in tilapia fish at 500 mg/kg-1. Treated groups showed significantly better immune parameters compared to the control group (p < 0.05). Conclusions: C. vulgaris crude biomass in a fish meal at a lower incorporation level of 5% can increase specific and non-specific immunity in tilapia fish in a shorter time duration.

Keywords: Chlorella vulgaris, hematology profile, immune boost, lymphoproliferation

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